Immuno-detection of the virulence determinant OmpX at the cell surface of Enterobacter cloacae.
A model for the topology of the Enterobacter cloacae outer membrane protein OmpX has been proposed, based on the primary sequence and on analogy to homologous proteins. According to this model the membrane embedded part of the protein consists of eight antiparallel beta-strands. Four random coil loops are located at the bacterial surface and three beta-turns at the periplasmic side of the membrane. Antibodies were raised against synthetic peptides representing five OmpX domains, four of which are putative peripheral and one located in the membrane. The accessibilities of OmpX to these antibodies were tested in intact cells by immuno-gold electron microscopy. This study showed that OmpX is indeed an outer membrane protein, the N-proximal loop of which forms an IgG-accessible epitope at the cell surface.
de Kort
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G
Department of Medical Microbiology, Leiden University Medical Center, The Netherlands.
van der Bent-Klootwijk
P
P
van de Klundert
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D002851:Chromatography, High Pressure Liquid; D016972:Enterobacter cloacae / Q000737:chemistry / Q000235:genetics* / Q000472:pathogenicity; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D029968:Escherichia coli Proteins; D006867:Hydrolases; D007150:Immunohistochemistry; D008070:Lipopolysaccharides / Q000276:immunology; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis; D010957:Plasmids; D011817:Rabbits; D014774:Virulence
D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; D010455:Peptides; C482905:OmpX protein, E coli; D006867:Hydrolases